Nucleic acids for treatment of allergies

Abstract

The present invention provides DNA vaccines for the treatment or prevention of an allergic response. The vaccines comprise the coding sequence for Allergen X or fragments thereof fused in-frame with the lumenal domain of the lysosomal associated membrane protein (LAMP) and the targeting sequence of LAMP. The vaccines allow for presentation of properly configured three dimensional epitopes for production of an immune response when administered to a subject. The vaccines can be multivalent molecules, and/or can be provided as part of a multivalent vaccine containing two or more DNA constructs.

What is claimed: 
     
       1. An isolated nucleic acid encoding a polypeptide comprising:
 (a) a lysosomal associated membrane protein (LAMP) lumenal domain; 
 (b) at least one allergen comprising the amino acid sequence of SEQ ID NO: 45, 49, 53, 57, and/or 61 and 
 (c) a LAMP transmembrane domain/cytoplasmic tail. 
 
     
     
       2. The nucleic acid of  claim 1 , wherein LAMP is selected from LAMP polypeptide (LAMP-1), DC-LAMP, LAMP-2, LAMP-3, LIMP II, or ENDOLYN. 
     
     
       3. The nucleic acid of  claim 2 , wherein the LAMP lumenal domain is derived from LAMP-1. 
     
     
       4. The nucleic acid of  claim 3 , wherein the LAMP lumenal domain comprises an amino acid sequence chosen from:
 (a) SEQ ID NO:2; 
 (b) SEQ ID NO:3; 
 (c) amino acids 29-381 of SEQ ID NO:4 or 25-370 of SEQ ID NO:5; or 
 (d) a variant of any one of (a)-(c), wherein the variant comprises an amino acid sequence that is at least about 90%, at least about 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence of any one of (a)-(c). 
 
     
     
       5. The nucleic acid of  claim 1 , wherein the LAMP transmembrane domain/cytoplasmic tail comprises an amino acid sequence chosen from:
 (a) SEQ ID NO:1; 
 (b) amino acids 382-417 of SEQ ID NO:4 or 371-406 of SEQ ID NO:5; or 
 (c) a variant of (a) or (b), wherein the variant comprises an amino acid sequence that is at least about 90%, at least about 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence of (a) or (b). 
 
     
     
       6. The nucleic acid of  claim 1 , wherein the nucleic acid comprises:
 (a) any one of the polynucleotide sequences of SEQ ID NO: 44, 48, 52, 56 or 60; 
 (b) a polynucleotide encoding a polypeptide encoded by any one of the polynucleotide sequences of SEQ ID NO: 44, 48, 52, 56 or 60; 
 (c) a polynucleotide that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 45, 49, 53, 57 and/or 61; 
 (d) a polynucleotide encoding a LAMP lumenal domain one or more of SEQ ID NO: 45, 49, 53, 57, and/or 61, and a LAMP transmembrane domain/cytoplasmic tail; 
 (e) a polynucleotide encoding a LAMP lumenal domain and one or more of SEQ ID NO: 45, 49, 53, 57, and/or 61; or 
 (f) a polynucleotide encoding one or more of SEQ ID NO: 45, 49, 53, 57, and/or 61 and a trafficking domain LAMP transmembrane domain/cytoplasmic tail. 
 
     
     
       7. The nucleic acid of  claim 6 ,
 wherein the LAMP lumenal domain comprises:
 (a) SEQ ID NO:2; 
 (b) SEQ ID NO:3; 
 (c) SEQ ID NO:4 or SEQ ID NO:5; or 
 (d) a variant of any one of (a)-(c), wherein the variant comprises an amino acid sequence that is at least about 90%, at least about 95%, 96%, 97%, 98% or 99% identical to any one of the sequences shown in (a)-(c); 
 
 and wherein the LAMP transmembrane domain/cytoplasmic tail comprises:
 (a) SEQ ID NO:1; 
 (b) SEQ ID NO:4 or SEQ ID NO:5; or 
 (c) a variant of (a) or (b), wherein the variant comprises an amino acid sequence that is at least about 90%, at least about 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence of (a) or (b). 
 
 
     
     
       8. The nucleic acid of  claim 1 , wherein the nucleic acid encodes the amino acid sequence of SEQ ID No: 45. 
     
     
       9. The nucleic acid of  claim 3 , wherein the LAMP lumenal domain comprises:
 (a) SEQ ID NO:2 or 
 (b) a variant of (a), wherein the variant comprises an amino acid sequence that is at least about 90%, at least about 95%, 96%, 97%, 98% or 99% identical to (a). 
 
     
     
       10. The nucleic acid of  claim 3 , wherein the LAMP transmembrane domain/cytoplasmic tail comprises:
 (a) SEQ ID NO:1; or 
 (b) a variant of (a), wherein the variant comprises an amino acid sequence that is at least about 90%, at least about 95%, 96%, 97%, 98% or 99% identical to (a). 
 
     
     
       11. A vector comprising the nucleic acid of  claim 1 . 
     
     
       12. A host cell comprising the nucleic acid of  claim 1  or comprising a vector comprising the nucleic acid. 
     
     
       13. A vaccine comprising (a) the nucleic acid of  claim 1 , (b) a vector comprising the nucleic acid, or (c) a host cell comprising either the nucleic acid or the vector. 
     
     
       14. A method of treating or preventing an allergic response, wherein the method comprises administering to a subject the vaccine of  claim 13  in an effective amount sufficient to reduce, inhibit or prevent an allergic response. 
     
     
       15. The method of  claim 14 , wherein the vaccine is administered therapeutically. 
     
     
       16. The method of  claim 14 , wherein the vaccine is administered prophylactically. 
     
     
       17. The method of  claim 14 , wherein the method comprises a priming step and at least one boosting step. 
     
     
       18. The method of  claim 17 , wherein:
 (a) nucleic acid, vector, or host cell is administered to the subject in the priming step; and/or 
 (b) the nucleic acid, vector, or host cell is administered to the subject in the boosting step. 
 
     
     
       19. The method of  claim 17 , wherein the allergen used to prime is the same that is used to boost. 
     
     
       20. The method of  claim 17 , wherein the allergen used to prime is derived from the same protein as the allergen used to boost. 
     
     
       21. The method of  claim 17 , wherein more than one allergen is used to prime and/or boost.